Feuillassier Lionel, Masanet P., Romans P., Barthelemy D., Engelmann Florent. (2015). Towards a vitrification-based cryopreservation protocol for the coral Pocillopora damicornis L. : tolerance of tissue balls to 4.5 M cryoprotectant solutions. Cryobiology, 71 (2), p. 224-235. ISSN 0011-2240.
Titre du document
Towards a vitrification-based cryopreservation protocol for the coral Pocillopora damicornis L. : tolerance of tissue balls to 4.5 M cryoprotectant solutions
Feuillassier Lionel, Masanet P., Romans P., Barthelemy D., Engelmann Florent
Source
Cryobiology, 2015,
71 (2), p. 224-235 ISSN 0011-2240
In this study, we tested the tolerance of tissue balls (TBs, 100-400 mu m in diameter) from the coral Pocillopora damicornis produced using mechanical excision to exposure to cryoprotectant (CPA) solutions. TBs were treated for 20 mm at room temperature with individual, binary, ternary or quaternary CPA solutions with a total molarity from 2.0 to 5.0 M. Four CPAs were used: ethylene glycol (EG), dimethylsulfoxide (Me2SO), methanol (Met) and glycerol (Gly). In some experiments, the molarity of the CPA solutions was increased and decreased in a stepwise manner. The tolerance of TBs following CPA treatment was evaluated using two parameters. The Tissue Ball Regression (expressed in mu m/h) measured the diameter regression of TBs over time. The % Undamaged TBs quantified the proportion of TBs, which remained intact over time after the CPA treatment. TBs tolerated exposure to binary solutions with a total molarity of 4.0 M containing 2.0 M EG + 2.0 M Met and 2.0 M EG + 2.0 M Gly. TBs displayed tolerance to ternary solutions with a total molarity up to 3.0 M, containing each CPA at 1.0 M. Quaternary solutions with a total molarity of 4.0 M containing each CPA at 1.0 M were not tolerated by TBs. When the molarity of the CPA solutions was increased and decreased in a stepwise manner, TBs withstood exposure to a CPA solution with a total molarity of 4.5 M, containing 1.5 M EG + 1.5 M Gly + 1.5 M Me2SO. This study confirmed the interest of using TBs to test CPA solutions, with the objective of developing a vitrification-based cryopreservation protocol.
